CsHD1 protein and gene related to apical development and dwarfing and application thereof

A protein and top technology, applied in application, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2021-07-23
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Citrus self-pruning is essentially a physiological process of active shedding of the top growth area of ​​shoots. Although this self-pruning process is of great significance in the growth and development of various plants, so far, there is no detailed report on it. The rationality of this phenomenon and a reasonable explanation in terms of evolution, molecular regulation mechanism, etc.

Method used

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  • CsHD1 protein and gene related to apical development and dwarfing and application thereof
  • CsHD1 protein and gene related to apical development and dwarfing and application thereof
  • CsHD1 protein and gene related to apical development and dwarfing and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Cloning of CsHD1 Gene

[0019] This embodiment provides a method for cloning the CsHD1 gene, and the specific steps are as follows:

[0020] (1) Extraction of plant RNA

[0021] The collection of plant tissue samples (sweet orange top tissue) needs to use ice boxes or liquid nitrogen, and immediately store them in a -80°C refrigerator for later use. The whole process of RNA extraction needs to be carried out in an ultra-clean workbench, and the reagent adopts the Trizol kit of Takara Company, and the main steps are as follows:

[0022] a. Take 1mL RNAiso Plus reagent and put it in a 2mL RNAase free centrifuge tube, put it on ice;

[0023] b. Liquid nitrogen sample grinding: take an appropriate amount of sample and place it in a mortar, pour liquid nitrogen into it, and quickly grind it to powder;

[0024] c. Take an appropriate amount of ground sample in the RNAiso Plus solution, add 1 / 5 volume of chloroform after standing at room temperature, oscillate and...

Embodiment 2

[0062] Embodiment 2 Transformation of Arabidopsis

[0063] The transformation process of Arabidopsis thaliana is as follows: when the Arabidopsis thaliana in the culture room grows up to flowering, the genetic transformation of Arabidopsis thaliana is carried out by the inflorescence infection method. First, the required Agrobacterium is shaken or streaked to activate, The activated bacterial liquid was used for the transformation experiment of Arabidopsis thaliana, and the specific operation steps were as follows:

[0064] a. Preparation of osmotic culture solution: 1 / 2MS solution or 5% sucrose solution, 0.5g of MES was added per liter, and the pH was adjusted to about 5.7;

[0065] b. The day before the transformation, prepare the overnight culture of Agrobacterium to the OD of the bacterial solution 600 Reach 1.2-1.6;

[0066] c. Centrifuge at room temperature at 5000rpm for 15min to collect the bacteria;

[0067] d. Discard the supernatant and suspend the cells with the...

Embodiment 3

[0072] Example 3: Genetic Transformation of Tobacco

[0073] Prepare the Agrobacterium bacteria solution one day in advance and culture overnight in a shaker at 28°C (OD 600 About 1.2), the next day, the cultured bacterial liquid was centrifuged to collect the bacterial cells, and the bacterial cells obtained by suspending the anti-MS liquid medium, OD 600 The value is adjusted to 0.8-1.0 for standby, and the whole transformation process is carried out in the ultra-clean workbench. The detailed transformation operation steps are as follows:

[0074] a. Take healthy and well-grown wild-type tobacco leaves and cut off the main veins, cut them into small pieces with a width of about 1 cm (appropriately make a small wound to facilitate Agrobacterium infection), and place them in the infection solution with the back facing up. About 10 minutes, during which it kept shaking;

[0075] b. Take out the leaves with sterile tweezers and place them on sterile filter paper to blot the re...

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Abstract

The invention relates to the technical field of plant genetic engineering, in particular to a CsHD1 protein and gene related to apical development, late flowering and dwarfing and application thereof. The CsHD1 gene is separated from sweet oranges for the first time and subjected to functional identification, and it is found that the CsHD1 gene can promote apical development of citrus, dwarf plants and prolong the vegetative growth period of the tops of transgenic plants. In addition, the gene enables transgenic plants to show phenotypes such as dwarfing and leaf crumpling, and can greatly prolong the vegetative growth stage of the tops of the plants so as to delay the flowering time. According to the invention, the regulation and control mechanism of apical development and dwarfing of citrus is analyzed, regulation and control and improvement of citrus plant types are facilitated in actual production, and the purpose of increasing both production and income is finally achieved by using genetic engineering means.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to a CsHD1 protein, gene and application thereof related to top development and dwarfing. Background technique [0002] Citrus is the largest fruit tree in the world. It is a perennial woody plant. During the annual growth cycle of citrus, three types of new shoots are usually produced, which grow in spring, summer and autumn. They are called spring shoots, spring shoots, and autumn shoots respectively. In summer shoots and autumn shoots, the tops of new shoots will fall off after they develop to a certain extent. This phenomenon is common in almost all citrus varieties, which is called the process of "self-cutting" of terminal buds. Citrus self-pruning is essentially a physiological process of active shedding of the top growth area of ​​shoots. Although this self-pruning process is of great significance in the growth and development of various plants, so far, ther...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/00A01H6/82A01H6/20A01H6/78
CPCC07K14/415C12N15/8261C12N15/8205
Inventor 张金智胡春根曾仁芳付黎明
Owner HUAZHONG AGRI UNIV
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